A new canthinone glycoside isolated from the root barks of Ailanthus altissima with NO inhibitory activity.

One new canthinone glycoside (1), together with six known compounds (2-7) including three lignans (2-4), two coumarins (5-6) and one phenol (7) was isolated from the root barks of Ailanthus altissima. The structure of new compound 1 was established by the interpretation of UV, IR, MS and NMR data, while its absolute configuration was determined by acid hydrolysis and GIAO NMR calculations with DP4+ probability analysis. The inhibitory effects of all compounds on Nitric oxide (NO) production were investigated in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Results showed that compounds 2 and 5 displayed NO production inhibitory activity with IC50 values of 30.1 and 15.3 µM, respectively.

In Vitro Screening of Ecotoxic and Cytotoxic Activities of Ailanthus altissima Leaf Extract against Target and Non-Target Plant and Animal Cells.

Ailanthus altissima, an invasive plant species, exhibits pharmacological properties, but also some allergic effects on humans. This study aimed to evaluate the potential toxicity of A. altissima leaves, using a complex approach towards different organisms. The ecotoxic impact of a crude extract was investigated on seeds germination and brine shrimp lethality. Cytotoxicity was studied in vitro using non-target (haemolysis, liposomal model, fibroblast), and target (cancer cells) assays. Leaf extract at 1000 µg/mL significantly inhibited wheat and tomato germination, while no significant effects were found on parsley germination. A slight stimulatory effect on wheat and tomato germination was found at 125 µg/mL. In a brine shrimp-test, the extract showed a low toxicity at 24 h post-exposure (LC50 = 951.04 ± 28.26 µg/mL), the toxic effects increasing with the exposure time and extract concentration. Leaf extract caused low hematotoxicity. The extract was biocompatible with human gingival fibroblasts. No anti-proliferative effect was found within the concentration range of 10-500 µg/mL on malignant melanoma (MeWo) and hepatocellular carcinoma (HepG2). In a liposomal model-test, the extract proved to possess low capability to alter the eukaryotic cell-mimicking membranes within the tested concentration range. Given the low to moderate toxicity on tested organisms/cells, the A. altissima autumn leaves may find useful applications.

Phytoremediation potential of seedlings: comparing heavy metal accumulation in Ailanthus, Acer, and Fraxinus species.

This study investigates the phytoremediation potential of non-productive seedlings of Ailanthus altissima, Acer pseudoplatanus, and Fraxinus excelsior for lead, cadmium, and zinc accumulation in contaminated soils of Zanjan Province, an industrial area with significant pollution. The evaluation employed a completely randomized design, with three treatment levels for each element, alongside a control treatment, replicated three times over a two-year period. A total of 810 one-year-old seedlings from the three species were involved in the study. Soil contamination levels, ranging from 0 to 2000 mg/kg for lead and zinc and from 0 to 200 mg/kg for cadmium, were administered through soil pot irrigation. Sampling of seedling stems and pot soils was conducted in November of 2021 and 2022. The absorption levels of elements in the samples were determined using the dry acid digestion method and an ICP-OES atomic absorption spectrometer. Results indicate species-specific variations in metal absorption, with Ailanthus showing the highest accumulation rates. Findings suggest Ailanthus as a promising candidate for soil improvement in polluted environments, particularly in contaminated soils of Zanjan Province.

Transcriptome analysis reveals the fruit color variation in Ailanthus altissima.

Anthocyanins are responsible for the intensity of color in plants; however, the systematic mechanisms underlying the color differences in the fruit of Ailanthus altissima remain unknown. Therefore, this study aims to analyze the transcriptomes of the white and red fruit of A. altissima by screening and validating the key genes involved in flavonoid and anthocyanin biosynthesis. Samples of A. altissima fruit were collected 30, 45, and 60 days after flowering, and their pigment and sugar content were determined. The anthocyanin content was significantly higher in red than in white fruits. Transcriptome analysis was also performed on the fruit samples, 73,807 unigenes were assembled and annotated to seven databases. Twenty-one co-expressed modules were identified via weighted gene co-expression network analysis, of which two were associated with flavonoids and anthocyanins. Furthermore, in three growth stages, 126, 30, and 124 differentially expressed genes were screened between white and red fruit. Genes involved in flavonoid and anthocyanin metabolism were identified. AaDFR (A. altissima bifunctional dihydroflavonol 4-reductase/flavanone 4-reductase) and AaANS (A. altissima anthocyanidin synthase) were associated with flavonoid and anthocyanin metabolism. Members of the AaDFR and AaANS families were also identified, and their basic physicochemical characteristics, conserved domains, motif compositions, phylogenetics, and expression levels were analyzed. The overexpression of AaDFR and AaANS in transgenic Arabidopsis significantly increased the content of seed and foliar flavonoids and anthocyanins. The study elucidated the different mechanisms underlying fruit color development and provided insight into A. altissima plants breeding with commercially desirable properties.

Genome Resources of Verticillium dahliae VdGL16: The Causal Agent of Vascular Wilt on the Invasive Species Ailanthus altissima.

Verticillium species are known as plant pathogens responsible for wilt diseases in a large variety of dicotyledon plants and crops in many parts of the world. Here we present the draft genome sequence of Verticillium dahliae Kleb. (strain VdGL16) isolated in Italy from the invasive alien species Ailanthus altissima (Mill.; commonly known as tree-of-heaven) showing Verticillium wilt symptoms. The comparison between the newly sequenced genome with those publicly available revealed candidate genes putatively involved in pathogenicity. The genome represents a new useful source for future research on Verticillium genetics and biology as well as research on novel approaches in the control of A. altissima.

A putative new emaravirus isolated from Ailanthus altissima (Mill.) Swingle with severe crinkle symptoms in China.

A putative new emaravirus, named "ailanthus crinkle leaf-associated emaravirus" (ACrLaV), was detected in Ailanthus altissima with severe crinkle symptoms by RNA-Seq and RT-PCR. Four viral segments associated with ACrLaV were identified and fully sequenced, except for a few nucleotides at the genomic termini. The RNA-dependent RNA polymerase (RNA1), glycoprotein (RNA2), nucleocapsid protein (RNA3), and movement protein (RNA4), showed 26.5%-57%, 17%-49.9%, 14.4%-40.4%, and 14.1%-65.9% amino acid sequence identity, respectively, to those of known emaraviruses. All four ACrLaV genomic RNA segments are most closely related to those of common oak ringspot-associated virus from Germany, as supported by sequence comparisons and phylogenetic analysis. ACrLaV is considered a distinct member of the genus Emaravirus, and this is the first report of an emaravirus in A. altissima.

Transcriptome sequencing and microsatellite marker discovery in Ailanthus altissima (Mill.) Swingle (Simaroubaceae).

Ailanthus altissima Swingle, is a tree species native to East Asia and has a great potential in decorative, bioenergy and industrial applications in many countries. To date, despite its commercial importance, the genomic and genetic resources available for this species are still insufficient. In this study, we characterized the transcriptome of A. altissima and developed thirteen EST-SSRs (expressed sequence tag-simple sequence repeats) based on Illumina paired-end RNA sequencing (RNA-seq). Besides, we developed ten polymorphic chloroplast microsatellite (cpSSR) markers using the available chloroplast genome of A. altissima. The transcriptome data produced 87,797 unigenes, of which 64,891 (73.91%) unigenes were successfully annotated in at least one protein database. For cpSSR markers the number of detected alleles (N) per marker varied from three at cpSSR12 to twelve at cpSSR8, the unbiased haploid diversity indices (uh) varied from 0.111 to 0.485, and haploid diversity indices (h) ranged from 0.101 to 0.444 with an average unbiased haploid diversity index (uh) of 0.274. Overall, a total of 65 different cpSSR alleles were identified at the ten loci among 165 individuals of A. altissima. The allele number per locus for EST-SSRs varied from 2.143 to 9.357, and the values of observed and expected heterozygosity ranged from 0.312 to 1.000 and 0.505 to 0.826, respectively. The molecular markers developed in this study will facilitate future genetic diversity, population structure, long distance-gene transfer and pollen-based gene flow analyses of A. altissima populations from its known distribution ranges in China focusing on planted and natural forest stands.

Benzoic acid derivatives from the root barks of Ailanthus altissima.

Two new benzoic acid derivatives (1-2), together with four known compounds (3-6) have been isolated from the n-BuOH soluble fraction of ethanolic extract from Ailanthus altissima. The gross structures of the new compounds were deduced by detailed spectroscopic analysis including HRESIMS and 1D/2D NMR spectroscopy. The stereochemistry of 1 was determined by modified Mosher's method. All compounds were evaluated for their neuroprotective effects against H2O2-induced oxidative stress in human neuroblastoma SH-SY5Y cells and none of them displayed obvious neuroprotective activities. [Formula: see text].

Structure elucidation of a new terpenylated coumarin with the combination of CASE algorithms and DFT/NMR approach.

One new terpenylated coumarin ailanthuscoumarin was isolated from the root barks of Ailanthus altissima (Mill.) Swingle. The high oxidation of the compound led to the difficulty of structure elucidation by 2D-NMR spectra. Its structure was determined with the aid of computer-assisted structure elucidation (CASE) expert systems and Gauge-independent atomic orbital (GIAO) NMR calculations. The absolute configuration of ailanthuscoumarin was determined by the comparison between the experimental and calculated electronic circular dichroism (ECD) spectra. The anti-hepatoma activity of ailanthuscoumarin against two human hepatoma cells (Hep3B, HepG2) was also reported.

Effects of Ailanthus altissima Invasion and Removal on High-Biodiversity Mediterranean Grasslands.

Ailanthus altissima is one of the worst invasive plants in Europe in several habitat types, including high-biodiversity grasslands. The aim of this work was to evaluate the impact of the invasive A. altissima on high-biodiversity grassland vegetation and the effects of its removal on the recovery of native plant communities. The study area was within the Alta Murgia National Park (SE Italy). Seventeen vegetation quadrats were sampled in invaded grasslands and nine quadrats were sampled in nearby uninvaded areas. A. altissima was removed from six quadrats, which were sampled for two years after plant removal. Cluster analysis and non-metric multidimensional scaling ordination were used to identify and visualize the general vegetation pattern. Generalised Linear Models with different error structures were used to analyse the effects of A. altissima on native grasslands and vegetation recovery after removal. Results showed that the invasion of A. altissima changed drastically the community composition, reduced plant richness and diversity. Invaded stands had a greater presence of ruderal and widely distributed taxa, as opposed to a lesser presence of endemic and Mediterranean ones. The differences in the community composition between invaded and uninvaded quadrats became clearly detectable when A. altissima plants exceeded a threshold of 1 m of height and 50% of coverage. After A. altissima removal, the recovery of the grassland community was not completely achieved after two years.

Anticancer properties and mechanism of action of the quassinoid ailanthone.

Ailanthone (AIT) is a quassinoid natural product isolated from the worldwide-distributed plant Ailanthus altissima. The drug displays multiple pharmacological properties, in particular significant antitumor effects against a variety of cancer cell lines in vitro. Potent in vivo activities have been evidenced in mice bearing hepatocellular carcinoma, nonsmall cell lung cancer and castration-resistant prostate cancer. This review focusses on the mechanism of action of AIT, notably to highlight the capacity of the drug to activate DNA damage responses, to inhibit the Hsp90 co-chaperone p23 and to modulate the expression of several microRNA. The interconnexion between these effects is discussed. The unique capacity of AIT to downregulate oncogenic miR-21 and to upregulate the tumor suppressor miRNAs miR-126, miR-148a, miR-195, and miR-449a is presented. AIT exploits several microRNAs to exert its anticancer effects in distinct tumor types. AIT is one of the rare antitumor natural products that binds to and strongly inhibits cochaperone p23, opening interesting perspectives to treat cancers. However, the toxicity profile of the molecule may limit its development as an anticancer drug, unless it can be properly formulated to prevent AIT-induced gastro-intestinal damages in particular. The antitumor properties of AIT and analogs are underlined, with the aim to encourage further pharmacological studies with this underexplored natural product and related quassinoids. HIGHLIGHTS: Ailanthone (AIT) is an anticancer quassinoid isolated from Ailanthus altissima It inhibits proliferation and induces cell death of many cancer cell types The drug activates DNA damage response and targets p23 cochaperone Up or downregulation of several microRNA by AIT contributes to the anticancer activity Analogs or specific formulations must be developed to prevent the toxicity of AIT.

The Protective Role of Bark and Bark Fibers of the Giant Sequoia (Sequoiadendron giganteum) during High-Energy Impacts.

The influences of (1) a high fiber content, (2) the arrangement of fibers in fiber groups, and (3) a layered hierarchical composition of the bark of the giant sequoia (Sequoiadendron giganteum) on its energy dissipation capability are analyzed and discussed regarding the relevance for an application in bioinspired components in civil engineering. The giant sequoia is native to the Sierra Nevada (USA), a region with regular rockfalls. It is thus regularly exposed to high-energy impacts, with its bark playing a major protective role, as can be seen in the wild and has been proven in laboratory experiments. The authors quantify the fundamental biomechanical properties of the bark at various length scales, taking into account its hierarchical setup ranging from the integral level (whole bark) down to single bark fibers. Microtensile tests on single fibers and fiber pairs give insights into the properties of single fibers as well as the benefits of the strong longitudinal interconnection between single fibers arranged in pairs. Going beyond the level of single fibers or fiber pairs, towards the integral level, quasistatic compression tests and dynamic impact tests are performed on samples comprising the whole bark (inner and outer bark). These tests elucidate the deformation behavior under quasistatic compression and dynamic impact relevant for the high energy dissipation and impact-damping behavior of the bark. The remarkable energy dissipation capability of the bark at the abovementioned hierarchical levels are linked to the layered and fibrous structure of the bark structurally analyzed by thin sections and SEM and µCT scans.

Two Novel Quassinoid Glycosides with Antiviral Activity from the Samara of Ailanthus altissima.

Phytochemistry investigations on Ailanthus altissima (Mill.) Swingle, a Simaroubaceae plant that is recognized as a traditional herbal medicine, have afforded various natural products, among which C20 quassinoid is the most attractive for their significant and diverse pharmacological and biological activities. Our continuous study has led to the isolation of two novel quassinoid glycosides, named chuglycosides J and K, together with fourteen known lignans from the samara of A. altissima. The new structures were elucidated based on comprehensive spectra data analysis. All of the compounds were evaluated for their anti-tobacco mosaic virus activity, among which chuglycosides J and K exhibited inhibitory effects against the virus multiplication with half maximal inhibitory concentration (IC50) values of 56.21 ± 1.86 and 137.74 ± 3.57 µM, respectively.

IgE-mediated allergic responses associated to Ailanthus altissima pollen using an animal model.

BACKGROUND: Murine models have been widely used in the study of allergy as sensitized mice can produce IgE and/or IgG1in response after the injection of an antigen/adjuvant combination. Ailanthus altissima pollen (AAP) has been recently reported as an emerging aeroallergen in Iran. So far, several AAP candidate allergens by the screening of allergen-specific IgE in the sera from AAP sensitized patients in Iran. OBJECTIVE: The aim of the present study was to detect and compare the allergens eliciting an IgE response in a mouse model, and in human, using pollen extract of A. altissima and an immunoproteomics based approach. METHODS: The pollen proteins were extracted in phosphate-buffered saline (PBS). Thirty male BALB/c mice were randomly divided into two groups of AP extract sensitized and sham that respectively received AAP PBS extract and a PBS control by intraperitoneal injections at regular intervals. The optimized AAP protein extracts were analyzed using 2D-gel electrophoresis and were subsequently confronted to pooled sera of sensitized mice. RESULTS: Two-D gel electrophoresis of AAP extract allowed the separation of 125 protein spots distributed in a wide range of pI and molecular masses. Two-DE immunoblotting using pooled sera of sensitized mice led to the detection of 14 IgE reactive spots with molecular masses ranging from 12 to 40-42kDa. CONCLUSION: The results do not correlate with our previous analyses using human AAP-sensitized sera. These findings reflect some differences in the sIgE reactivity to allergenic proteins in animal models.

Bioactive Polyphenols Modulate Enzymes Involved in Grapevine Pathogenesis and Chitinase Activity at Increasing Complexity Levels.

The reduction of synthetic chemistry use in modern viticulture relies on either the biological control of microorganisms or the induction of pathogenesis-related proteins. In the present study, the effects of hydro-alcoholic plant extracts (PEs) (i.e., by-products of Vitis vinifera L., leaves of Olea europaea L. and Ailanthus altissima (Mill.) Swingle) were tested on purified enzymes activity involved in plant-pathogen interactions. The polyphenolic composition was assayed and analyzed to characterize the extract profiles. In addition, suspension cell cultures of grapevine were treated with PEs to study their modulation of chitinase activity. Application of grape marc's PE enhanced chitinase activity at 4 g L-1. Additionally, foliar treatment of grape marc's PE at two doses (4 g L-1 and 800 g L-1) on grapevine cuttings induced a concentration-dependent stimulation of chitinase activity. The obtained results showed that the application of bioactive compounds based on PEs, rich in phenolic compounds, was effective both at in vitro and ex/in vivo level. The overall effects of PEs on plant-pathogen interaction were further discussed by applying a multi-criteria decision analysis, showing that grape marc was the most effective extract.

Year-to-year variation of the elemental and allergenic contents of Ailanthus altissima pollen grains: an allergomic study.

The Ailanthus altissima pollen (AAP) has been reported as an emerging aeroallergen worldwide. This paper aims at examining the allergen pattern and the elemental composition of A. altissima pollen collected during two consecutive seasons (2014 and 2015). A gel-based allergomic study and SEM coupled to energy-dispersive X-ray (EDX) analysis have been carried out in order to evaluate the allergenic and elemental composition of AAP in two consecutive years. The IgE reactive patterns of 2014 and 2015 AAP PBS extracts were compared using the serum of a 31-year-old woman suffering from severe pollinosis symptoms to AAP. The EDX analysis revealed an important year-to-year variation in the ratios of some polluting elements such as nickel, sulfur, aluminum, lead, and copper. Gel alignments and comparative immunoproteomic analyses showed differential protein expression and IgE reactive patterns between AAPs collected in 2014 and 2015 pollinating seasons. From 20 distinct IgE-reactive spots detected in AAP extracts, 13 proteins showed higher expression in 2014 sample, while 7 allergen candidates exhibited an increased expression in AAP collected in 2015. Matrix-assisted laser desorption ionization-MS/MS analyses led to the identification of 13 IgE-binding proteins with confidence, all belonging to well-known allergenic protein families, i.e., enolase, calreticulin, and pectate lyase. Overall, the 2014 AAP showed higher concentrations of urban polluting elements as well as an increased expression of allergenic pectate lyase isoforms of about 52 kDa. This study demonstrates that the implementation of allergomic tools for the safety assessment of newly introduced and invasive plant species would help to the comprehensive monitoring of proteomic and transcriptomic alterations involving environmental allergens.

Structurally Diverse Highly Oxygenated Triterpenoids from the Roots of Ailanthus altissima and Their Cytotoxicity.

Ten new triterpenoids, ailanaltiolides A-J (1-10), and three known analogues (11-13) were isolated from the roots of Ailanthus altissima. Compounds 1-7 are apotirucallane-type, compounds 8 and 9 are tirucallane-type, and compound 10 is a trinordammarane-type triterpenoid. This is the first study indicating the genus Ailanthus as a potential source for apotirucallane derivatives, which contain an α,ß-unsaturated-ε-lactone A-ring and diversely modified C-17 side chains. Spectroscopic data interpretation, electronic circular dichroism analysis, and X-ray crystallographic data defined the structures and absolute configurations of these triterpenoids. Compounds 2, 7, and 8 showed cytotoxicity against four tumor cell lines (HeLa, 786-O, HepG2, and A549). In particular, compound 2 exhibited the highest activity against 786-O cells with an IC50 value of 8.2 µM in vitro.

Anti-inflammatory Effects of Canthin-6-one Alkaloids from Ailanthus altissima.

Canthin-6-one (CO) alkaloids possess various biological activities, including antibacterial, antitumor, antifungal, and antiviral activities. However, their anti-inflammatory effects and underlying molecular mechanisms are poorly characterized. This study aimed to investigate the anti-inflammatory effects of CO and its derivative 5-(1-hydroxyethyl)-canthin-6-one (5-HCO), isolated from the stem barks of Ailanthus altissima in lipopolysaccharide (LPS)-stimulated macrophages. CO (1 and 5 µM) and 5-HCO (7.5 and 15 µM) significantly inhibited the LPS-induced expression of inducible nitric oxide synthase. In addition, CO (1 and 5 µM) and 5-HCO (15 µM) markedly suppressed the production of prostaglandin E2 (PGE2) and expression of cyclooxygenase-2, a key enzyme in PGE2 synthesis, in LPS-stimulated macrophages. Moreover, CO treatment significantly reduced monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-α (TNF-α) expression, whereas 5-HCO inhibited MCP-1, but not TNF-α expression. Both CO and 5-HCO inhibited the phosphorylation of inhibitor kappa B and transcriptional activation of nuclear factor kappa B (NF-κB) in LPS-stimulated macrophages. In addition, CO, but not 5-HCO, markedly reduced Akt phosphorylation. Taken together, these data suggest that CO, but not 5-HCO with a hydroxyethyl moiety on the D ring, has potent anti-inflammatory activity in LPS-stimulated macrophages through the downregulation of both the NF-κB and the Akt pathway.

Chipping to Destroy Egg Masses of the Spotted Lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae).

A chipping study was conducted during the winter of 2015 in Berks County, Pennsylvania, to determine efficacy against field collected egg masses of the spotted lanternfly, Lycorma delicatula (White) (Hemiptera: Fulgoridae). Infested Ailanthus altissima Swingle (Sapindales: Simaroubaceae) trees in eastern Pennsylvania were felled and egg masses were counted. Sections were either chipped or allowed to remain intact as controls. Chipped material and intact wood controls were placed in screened barrels and monitored for emergence. No L. delicatula nymphs were found in the chipped treatment, as opposed to hundreds of nymphs per barrel in the intact control treatment. We conclude that mid-winter chipping, using the standard 1-inch in 2-dimension chip size, is a quarantine safe mitigation method suitable for treating wood infested with L. delicatula egg masses.

The Complete Chloroplast Genome Sequence of Tree of Heaven (Ailanthus altissima (Mill.) (Sapindales: Simaroubaceae), an Important Pantropical Tree.

Ailanthus altissima (Mill.) Swingle (Simaroubaceae) is a deciduous tree widely distributed throughout temperate regions in China, hence suitable for genetic diversity and evolutionary studies. Previous studies in A. altissima have mainly focused on its biological activities, genetic diversity and genetic structure. However, until now there is no published report regarding genome of this plant species or Simaroubaceae family. Therefore, in this paper, we first characterized A. altissima complete chloroplast genome sequence. The tree of heaven chloroplast genome was found to be a circular molecule 160,815 base pairs (bp) in size and possess a quadripartite structure. The A. altissima chloroplast genome contains 113 unique genes of which 79 and 30 are protein coding and transfer RNA (tRNA) genes respectively and also 4 ribosomal RNA genes (rRNA) with overall GC content of 37.6%. Microsatellite marker detection identified A/T mononucleotides as majority SSRs in all the seven analyzed genomes. Repeat analyses of seven Sapindales revealed a total of 49 repeats in A. altissima, Rhus chinensis, Dodonaea viscosa, Leitneria floridana, while Azadirachta indica, Boswellia sacra, and Citrus aurantiifolia had a total of 48 repeats. The phylogenetic analysis using protein coding genes revealed that A. altissima is a sister to Leitneria floridana and also suggested that Simaroubaceae is a sister to Rutaceae family. The genome information reported here could be further applied for evolution and invasion, population genetics, and molecular studies in this plant species and family.